Esculetin ameliorates the lipopolysaccharides-induced infl ammation in neuroblastoma cells via the AMPKNLRP3 pathway

12^th International Conference on Stroke, Neurology and Cerebrovascular Diseases

August 18-19, 2021 | Webinar

Ziaur Rahman

Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research, Hyderabad, India 500037

Scientific Tracks Abstracts: JND

Abstract :

Aim: To investigate the effect of esculetin in lipopolysaccharide (LPS) induced neuroinfl ammation in Neuro-2a cells. Background: Since most of the available treatments for neuropathic pain are ineffective or produce unwanted effects, alternative therapeutic approaches are being investigated. This study evaluated the impact of esculetin on LPS-triggered infl ammatory cascades using Neuro-2a neuroblastoma cells. We hypothesized that esculetin might produce an anti-infl ammatory effect via the AMPKNLRP3 pathway. Methodology: The anti-infl ammatory and antioxidant effects of esculetin (10 & 20 μM) were assessed using the Neuro-2a neuroblastoma cells. The Neuro-2a cells were pre-treated with esculetin for 24 h and then exposed to the LPS (1 μg/mL) for one h. Following LPS incubation, 2',7'-dichlorodihydrofl uorescein diacetate (DCFDA), immunoblotting and immunocytochemistry assays were performed to examine the protective effect of esculetin. To confi rm the therapeutic effect of esculetin in neuropathic pain, a chronic constriction injury (CCI) model was developed in Sprague-Dawley rats. Results: As expected, treatment with LPS signifi cantly increased reactive oxygen species (ROS) production compared to control. Esculetin at 20 μM concentration signifi cantly reversed the increased levels of ROS. LPS-exposed Neuro-2a cells also showed an increased expression of nucleotide-binding oligomerization domain leucine-rich repeat and pyrin domain-containing (NLRP3), and decreased expression of nuclear factor erythroid 2-related factor-2 (Nrf-2) and 5' adenosine monophosphateactivated protein kinase-3 (AMPK-3) compared to the control group. Esculetin at both doses (10 & 20 μM) signifi cantly downregulated the expression of NLRP3 and increased the expression of Nrf-2 and AMPK-3 compared to the LPS-treated group. These results indicated the antioxidant and anti-infl ammatory effect of esculetin in LPS-induced neuroinfl ammation in Neuro-2a cells. In a preclinical study, the decreased pain threshold was seen in the paw withdrawal latency (sec), paw withdrawal threshold (g), paw withdrawal pressure (g), allodynia score, sciatic function index and proportion of postural index values in CCI animals compared to the sham group. Conclusion: Our results indicate the anti-infl ammatory and protective effects of esculetin via the AMPK-NLRP3 pathway. We also suggest that esculetin may reverse the CCI-induced lowered pain threshold.

Biography :

Ziaur Rahman is pursuing his PhD in Biological science under the supervision of Dr Manoj Dandaker. He completed his master in Pharmacology and Toxicology from NIPER, Mohali, India. He has published more than fi ve papers in reputed journals and serving as an editorial board member of the Biological Sciences journal. (Up to 100 words)